US20040142069A1 - Silage additive and a process for preparing silage using it - Google Patents

Silage additive and a process for preparing silage using it Download PDF

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US20040142069A1
US20040142069A1 US10/751,462 US75146204A US2004142069A1 US 20040142069 A1 US20040142069 A1 US 20040142069A1 US 75146204 A US75146204 A US 75146204A US 2004142069 A1 US2004142069 A1 US 2004142069A1
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silage
mother liquor
liquid
liquor
mother
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US10/751,462
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Motoharu Takeda
Makoto Wakabayashi
Masakazu Gotou
Akira Tsukahara
Kouji Yumura
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Ajinomoto Co Inc
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Ajinomoto Co Inc
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Assigned to AJINOMOTO CO., INC. reassignment AJINOMOTO CO., INC. ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: GOTOU, MASAKAZU, TAKEDA, MOTOHARU, TSUKAHARA, AKIRA, WAKABAYASHI, MAKOTO, YUMURA, KOUJI
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
    • A23K30/10—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder
    • A23K30/15—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging
    • A23K30/18—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging using microorganisms or enzymes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K10/00—Animal feeding-stuffs
    • A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/142—Amino acids; Derivatives thereof
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K20/00—Accessory food factors for animal feeding-stuffs
    • A23K20/10—Organic substances
    • A23K20/163—Sugars; Polysaccharides
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23K—FODDER
    • A23K30/00—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs
    • A23K30/10—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder
    • A23K30/15—Processes specially adapted for preservation of materials in order to produce animal feeding-stuffs of green fodder using chemicals or microorganisms for ensilaging

Definitions

  • the present invention relates to a silage additive which is effective for livestock animals and which makes lactic acid bacteria involving in the silage fermentation proliferate, makes it possible to save without spoiling the quality of silage raw material for a long period of term, depress aerobic deterioration of silage, improves the digestibility of a raw material and has an effect to impart the nitrogen component into the silage, and to a process for preparing a silage using it.
  • silage raw material is placed in an anaerobic state by shutting air out whereby a lactic acid fermentation is induced and the pH is rapidly lowered to depress growth of aerobic bacteria at the initial stage of silage fermentation and the subsequent butyric acid fermentation which is the initial stage of the anaerobic fermentation thereby preserving the nutritional value and palatability of the feed.
  • the ensilaging process has been commonly conducted in the livestock farms as a means to store or preserve feed raw material for ruminant, pig and poultry, and to improve its quality.
  • acid-treatment such as formic acid, ammonium formate, citric acid and the like.
  • various kinds of lactic acid bacteria preparations have been added as the starter for the proliferation, and molasses, glucose or the like have been added as a saccharide source.
  • silage lactic acid fermentation has almost no ability to decompose the fiber component contained richly in plant raw material and accordingly for the purpose of improving the digestibility of the raw material an enzyme preparation, urea, ammonia and so on have been added singly or in admixture of them.
  • Lactic acid fermentation which is the most important process for the quality of silage, is originally accomplished by naturally occurred lactic acid bacteria without adding any lactic acid bacteria preparation which is costly, since usually about 10 3 lactic acid bacteria is adhered to natural raw material.
  • the condition under which silage is prepared is various and the relationship of dominance is also various among and between lactic acid bacteria microflora and harmful fermentation bacteria microflora in the course of silage fermentation, and therefore there has been required a process by which lactic acid fermentation may be strongly promoted under any ensiling condition.
  • Roughages such as various kinds of grasses, forage crops such as barley, corn, common millet, feed rice and the like, food by-products such as tofu cake, sweet potato or potato starch pulp, liquor by-product and the like, remainder discharged from food service industry and food distribution industry, etc. have poor digestibility in most cases due to fibrous material, and therefore it is very important from the nutritional point of view to improve the digestibility of fibrous components.
  • ammonia, urea an expensive enzyme capable of decomposing fiber and so on are commercially available.
  • ammonia treatment involves dangerous operation. There has been required development of such an additive that it has effects to improve the quality and digestibility of silage and it is cheap, easy to handle and not dangerous.
  • An object of the present invention is to provide a silage additive for improving the quality of silage which is effective for livestock animals and which makes lactic acid bacteria in the silage fermentation proliferate, makes it possible to preserve to make good quality silage for a long period of term, depress aerobic deterioration of silage in order to prepare silage having a high lactic acid content and a high digestibility and being rich in the amount of nitrogen component.
  • Another object of the present invention is to provide a preparing process of a silage having a high feed value with an improved digestibility using said silage additive.
  • the first of the present invention is a silage additive which comprises a liquid by-product from an amino acid fermentation broth, said liquid by-product being adjusted to a pH in the range of from 3.0 to 6.0.
  • the second of the present invention is the silage additive wherein the liquid by-product from an amino acid fermentation broth is one from L-glutamic acid, L-lysine or L-phenylalanine fermentation broth.
  • the third of the present invention is a process for preparing a silage which comprises adding a liquid by-product from an amino acid fermentation broth to a silage raw material in an amount of 1.0-10.0% based on fresh weight of the raw material, said liquid by-product being adjusted to a pH in the range of from 3.0 to 6.0 and subjecting the resultant mixture to anaerobic fermentation.
  • the fourth of the present invention is the process for preparing silage wherein lactic acid bacteria and a saccharide are further added to the silage raw material.
  • the fifth of the present invention is the process for preparing silage wherein the liquid by-product from an amino acid fermentation broth is one from L-glutamic acid, L-lysine or L-phenylalanine fermentation broth
  • the liquid by-product from an amino acid fermentation broth for use in the present invention is the one produced when an amino acid was removed from an amino acid fermentation broth by an appropriate separation procedure, said amino acid fermentation broth being obtained by culturing a an amino acid-producing microorganism in a culture medium containing various kinds of carbohydrates such as molasses, tapioca, corn and the like as a saccharide source and various kinds of ammonia nitrogen raw materials such as ammonia, ammonium sulfate and the like as a nitrogen source
  • the terms “liquid by-product” as defined herein means:
  • a mother liquid, its concentrate and the liquid produced when the mother liquid or its concentrate was subjected to desalting treatment said mother liquid being obtained when various kinds of amino acid fermentation broths wherein the amino acid moiety is an acidic amino acid such as glutamic acid or the like and a neutral amino acid such as phenylalanine, threonine, tryptophan or the like had been adjusted to an isoelectric pH of individual amino acids with a mineral acid such as sulfuric acid, hydrochloric acid or the like, whereupon said amino acid has crystallized out and said amino acid crystals have been separated from the broth by a solid-liquid separation procedure.
  • the amino acid moiety is an acidic amino acid such as glutamic acid or the like and a neutral amino acid such as phenylalanine, threonine, tryptophan or the like had been adjusted to an isoelectric pH of individual amino acids with a mineral acid such as sulfuric acid, hydrochloric acid or the like, whereupon said amino acid has crystallized out and said amino acid crystal
  • a liquid and its concentrate there may be taken a liquid and its concentrate, said liquid being produced when the microorganisms have been removed from the mother liquor, and further one produced when the liquid or its concentrate free from the microorganisms has been subjected to desalting treatment.
  • Glutamic acid by-products Condenced liquid
  • IFN Glutamic acid by-products
  • the pHs of these amino acid fermentation liquid by-products lie in an acidic side of 3.0-6.0 in all cases.
  • the liquid by-products are rich in nitrogen including volatile ammonia nitrogen, amino acid organic nitrogen and nitrogen derived from the fermentation bacteria in addition to an amino acid.
  • they are rich in minerals like sulfur or chlorine derived from the inorganic acid used for pH adjustment of the amino acid fermentation broth as mineral.
  • they contain as trace ingredients other minerals than described in the above, saccharides, organic acids, fermentation bacteria, etc. All of these ingredients are nutrients for silage fermentation microorganism and rumen microorganism.
  • silage raw materials there can be taken roughages such as various kinds of pasture grasses, rice straw and the like, concentrated feeds such as barley, corn, common millet, feed rice and the like, food by-products such as tofu cake, sweet potato or potato starch pulp, liquor by-product and the like, food process residue discharged from food service industry and food distribution industry, etc.
  • the amino acid fermentation liquid by-product is added to the silage raw material in an amount of 1.0-10.0% based on the fresh weight of the raw material, preferably the water content is adjusted to 60-70 wt. %. Saccharides and lactic acid bacteria may be further added depending on the necessity.
  • the resultant uniform mixture is ensilaged by a simple procedure that it is filled in a stationary or movable type of silo or otherwise in a silage bag made of a synthetic resin such as polyethylene, polypropylene or the like, deaerated, hermetically sealed and anaerobically fermented for about 40 days whereby silage raw material can be stored for a long period of term.
  • a synthetic resin such as polyethylene, polypropylene or the like
  • saccharides there may be used molasses and a reducing sugar (for example, maltose, lactose, cane sugar, trehalose, glucose or the like). It is adequate that the additional amount of saccharides is 2% or more based on the fresh weight of the raw material in considering the sugar content in the raw material.
  • a reducing sugar for example, maltose, lactose, cane sugar, trehalose, glucose or the like. It is adequate that the additional amount of saccharides is 2% or more based on the fresh weight of the raw material in considering the sugar content in the raw material.
  • a lactic acid bacteria preparation may be added as starter, if necessary.
  • the kind of bacterium which may be utilized as suitable lactic acid bacteria preparation includes lactobacillus plantarum, lactobacillus casei and the like. It is adequate that the additional amount of the lactic acid bacteria preparation is 0.05-1.0% based on the fresh weight of the raw material although it may be varied depending on the sugar content in the raw material.
  • the liquid by-product from amino acid fermentation broth used in Examples 1-7 and 9 is the one from L-phenylalanine fermentation broth which was registered as liquid by-product nitrogen fertilizer “PAL” (Reg. No. 74,220) and which has a pH of 4.5 and the total nitrogen content of 5.0 wt. % wherein the ammonia nitrogen content is 3.5 wt. % (hereinafter, referred to as “mother liquor”).
  • PAL liquid by-product nitrogen fertilizer
  • lactic acid bacteria Lactobacillus plantarum was cultured for 24 hours in a 804 liquid culture medium (10 ml) to prepare inoculum (pH 4.87).
  • each of the culture mediums prepared above was seeded with 200 ⁇ l of bacteria Lactobacillus plantarum pre-cultivated in 804 liquid culture medium (10 ml) using a 16 ⁇ test tube and liquid stationary culture was conducted in a thermostatic water tank maintained at 30° C.
  • Each silage was opened on 40th day after its preparation and samples were taken. Tenfold amounts of distilled water was added to the sample and the mixture was ground with a mixer and filtered out with gauze. The filtrate was subjected to centrifugal separation at a rotation speed of 15,000 rpm for 5 minutes at 5° C. Twofold amounts of 6% perchloric acid was added to the supernatant liquid and the mixture was stirred and then subjected to centrifugal separation under the same condition as in the above. The supernatant liquid was filtered out with a 45 ⁇ m filter and the amounts of lactic acid and volatile fatty acids (hereinafter, these acids will be referred as “organic acid” collectively) were measured by liquid chromatography.
  • the proportion of butyric acid in total organic acid is lower in the mother liquor-treated groups where the proportion of the mother liquor is 50 wt. % or more than in the control group. This result shows that the addition of such mother liquor causes effects of depressing a defective fermentation and of reducing dry matter loss rate.
  • the nitrogen component may be imparted to the silage by the addition of the mother liquor so that the content of crude proteins in the silage is increased as the proportion of the mother liquor added increases. This result is shown in FIG. 3.
  • each of the above culture mediums having different proportions of the mother liquor was seeded with 200 ⁇ l of silage yeast pre-cultivated in PYG liquid culture medium (10 ml) using a 16 ⁇ test tube and shaking culture (200 times reciprocation motions/min.) was conducted in a incubator maintained to 30° C.
  • Barley whole crop bottled silage was prepared and was opened on 40th day after its preparation. The ethanol contents were measured immediately after the opening of the silage and on 8th day after the opening. There were prepared a series of mixed solutions of the mother liquor and molasses with various proportions of 80:20 to 20:80 (weight ratio), and each of the mixed solutions was added to barley whole crop in an amount of 3% based on the fresh weight of barley whole crop. The resultant mixture was filled in a 1 L bottle (filling density of 500 g FW/L) to prepare barley whole crop silage.
  • FIG. 5 shows the result of the measurements conducted immediately after the opening of the silage and on the 8th day after opening for the ethanol contents in the control group and the mother liquor-treated group silages. Also, the increasing ratios of the ethanol content (after 8 days from the opening/immediately after the opening) in the control group and the mother liquor-treated groups are shown in FIG. 6.
  • Ethanol is inherently produced by yeast or hetero type lactic acid bacteria during the silage fermentation. Both are facultative anaerobic microorganisms. Although silage is in aerobic state after the opening, when a material which is rich in residual sugar (in some cases, some sugar remains in kernels after the opening of silage) was subjected to the silage fermentation as in this test, the above-described microorganisms still continue their activities for a while. It can be understood from the result of this test that as the proportion of the mother liquor added is high the activities of these microorganisms are inhibited more effectively.
  • FIG. 7 shows the result of the measurements conducted immediately after the opening of the silage and on the 8th day after opening for the organic acid contents in the control and the mother liquor-treated group silages. Also, the increasing ratios of the organic acid content (after the 8 days from the opening/immediately after the opening) in the control group and the mother liquor-treated groups are shown in FIG. 8.
  • the number of mould colony was measured in nutrient agar medium (comprising 5 g of peptone, 3 g of beef extract, 15 g of gelatin and 1,000 ml of distilled water, pH 6.8). The measurement was conducted for silage samples taken immediately after the opening of silage, on 2 nd and 6 th days after opening, respectively.
  • Glutamic acid fermentation broth was adjusted to isoelectric pH of said amino acid with sulfuric acid, the precipitated amino acid crystals was separated from the broth by a solid-liquid separation procedure whereby the obtained mother liquor was adjusted to a pH of around 5 with ammonia and subjected to desalting treatment and concentration.
  • Liquid by-product from lysine fermentation broth said liquid by-product having pH of 4.2 and total nitrogen content of 5.9 wt. % wherein the ammonia nitrogen content is 4.7 wt. % (hereinafter, referred to as “Lys mother liquor”) and being obtained from the lysine fermentation broth by the procedures shown below.
  • Lysine fermentation broth was adjusted to a pH of about 3 with sulfuric acid and then passed through a column packed with a strongly acidic cationic resin whereupon said amino acid was attached on the cationic resin, and the effluent was separated into the two fractions of bacterial cells and supernatant liquid by centrifugation separation.
  • the supernatant liquid was adjusted to pH of about 5 with an aqueous sodium hydroxide solution and then subjected to desalting treatment and concentration.
  • the resultant concentrate was mixed with the above bacteria cells.
  • Each of the silages was opened on 40th day after its preparation and a sample was taken.
  • the quantity of distilled water equal to 10 times the quantity of the sample was added to the sample, and the mixture was pulverized with a mixer and filtered out with gauze.
  • the filtrate was subjected to centrifugal separation at a rotation speed of 15,000 rpm for 5 minutes at 5° C.
  • Twofold amounts of 6% perchloric acid were added to the supernatant liquid.
  • the mixture was stirred and then subjected to centrifugal separation under the same condition as in the above.
  • the supernatant liquid was filtered out with a 45 ⁇ m filter to measure the pH and to determine volatile fatty acids using a liquid chromatography.
  • FIG. 11 shows the pHs of the respective silages of control group, Glu mother liquor-, Lys mother liquor- and Phe mother liquor-treated groups.
  • FIG. 12 shows total organic acid concentrations involving in the respective silages of the control group, the Glu mother liquor-, Lys mother liquor- and Phe mother liquor-treated groups.
  • FIG. 13 shows the proportions of lactic acid and acetic acid in total organic acid in the respective silages of the control group, the Glu mother liquor-, Lys mother liquor- and Phe mother liquor-treated groups.
  • Sweet potato starch is produced focusing on Kagoshima and Miyazaki prefectures in Japan. Sweet potato starch pulp which is by-product of the above starch was employed in this Example.
  • Table 10 shows the general constituent of sweet potato starch pulp employed in this example. TABLE 10 General constituent of sweet potato pulp Crude Crude Crude Moisture Protein Fiber Ash % pH % DM* 1 % DM % DM 72 9.1 1.8 18.6 8.7
  • lactic acid bacteria was further added to the control group and Phe mother liquor-treated group in an amount of 10 5 c.f.u/g F W based on the fresh weight of the resultant mixture.
  • Table 11 shows each of the control groups 1 and 2 and the mother liquor-treated groups 1 and 2. TABLE 11 Control Control Treated Treated Group 1 Group 2 Group 1 Group 2 Mother Not Not Added Added Liquor added added Lactic Not Added Not Added Acid Added Added Bacteria
  • Each silage was opened on about 40th day after its preparation and sample was taken.
  • the quantity of distilled water equal to 10 times the quantity of sample was added to sample, and the mixture was pulverized with a mixer and filtered out with gauze.
  • the filtrate was subjected to centrifugal separation at a rotation speed of 15,000 rpm for 5 minutes at 5° C.
  • Two fold amounts of 6% perchloric acid was added to the supernatant liquid.
  • the supernatant liquid was stirred and then subjected to centrifugal separation under the same condition as in the above.
  • the supernatant liquid was filtered out a 45 ⁇ m filter to measure the pH and to determine organic acids using a liquid chromatography.
  • FIG. 14 shows the pHs of the respective silages of the control groups and the mother liquor-treated groups.
  • FIG. 15 shows the proportions of each organic acid in total organic acid in the respective silages of the control groups and the mother liquor-treated groups.
  • FIG. 1 is a graph showing changes in the absorbance by culture time in the case where silage lactic acid bacteria Lactobacillus plantarum was cultured using mother liquor alone, molasses alone, mother liquor-molasses mixed solutions or glucose alone as silage additives.
  • FIG. 2 is a graph showing the proportions of lactic acid, acetic acid and butyric acid in total organic acid involving in the silage, said silage being prepared when barley whole crop bottled silage test has been carried out using the mother liquor-molasses mixed solution containing different amounts of the mother liquor as silage additives.
  • FIG. 3 is a graph showing changes in the content of crude proteins in the silage by the proportion of the mother liquor added, the silage being prepared when barley whole crop bottled silage test was carried out using the mother liquor-molasses mixed solution containing different amounts of the mother liquor added as silage additives.
  • FIG. 4 is a graph showing changes in the absorbance by culture time in the case where silage yeast was cultured using mother liquor alone, molasses alone, mother liquor-molasses mixed solutions or glucose alone as silage additives.
  • FIG. 5 is a graph showing the ethanol contents measured immediately after the opening of silage and on the 8 th day in the control group and the mother liquor-treated groups.
  • FIG. 6 is a graph showing the increasing ratios of the ethanol content (after 8 days from the opening of silage/immediately after the opening of silage) in the control group and the mother liquor-treated groups.
  • FIG. 7 is a graph showing the organic acid contents immediately after the opening of silage and on the 8 th day in the control group and the mother liquor-treated groups.
  • FIG. 8 is a graph showing the increasing ratio of the organic acid content (after 8 days from the opening of silage/immediately after the opening of silage) in the control group and the mother liquor-treated groups.
  • FIG. 9 is a graph showing the NDF contents in stem and leaf parts in the respective silages of the control group and the mother liquor-treated group.
  • FIG. 10 is a graph showing dry matter digestibility and NDF digestibility determined after immersion in the rumen of the cattle for 24 hours using in situ rumen digestibility technique for the respective silages of the control group and the mother liquor-treated groups.
  • FIG. 11 is a graph showing the pH of the silage prepared when Italian ryegrass bottled silage test has been carried out using each of the three kinds of amino acid fermentation broth liquid by-products as silage additives.
  • FIG. 12 is a graph showing the total organic acid concentrations in the control group and in the treatment groups of each of the three kinds of amino acid fermentation broth liquid by-products.
  • FIG. 13 is a graph showing the proportions of lactic acid and acetic acid in total organic acid in the control group and in the treatment groups of each of the three kinds of amino acid fermentation broth liquid by-products.
  • FIG. 14 is a graph showing the pH of the control group and the mother liquor-treated group silages prepared when sweet potato starch pulp bag silage test has been carried out.
  • FIG. 15 shows the proportions of individual organic acids in total organic acid in the control group and the mother liquor-treated group silages prepared when sweet potato starch pulp bag silage test has been carried out.

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CN109221684A (zh) * 2018-07-20 2019-01-18 马边下溪镇青山莲畜禽养殖专业合作社 一种饲草青储的方法
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CN114018912A (zh) * 2021-11-09 2022-02-08 江苏农牧科技职业学院 一种不同添加剂对黑麦草青贮品质影响的试验方法
CN114468123A (zh) * 2022-03-16 2022-05-13 四川水利职业技术学院 一种青贮饲料添加剂及其制备方法和应用
CN115886130A (zh) * 2022-12-08 2023-04-04 湖南农业大学 山银花在提高紫花苜蓿混合青贮饲料品质中的应用

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JP5224018B2 (ja) * 2006-05-29 2013-07-03 志戸本 宗徳 サイレージ及びその調製方法
DE602007013065D1 (de) 2006-07-14 2011-04-21 Legerth Jens Hoffner Homofermentierte produkte
WO2010150136A1 (en) 2009-06-22 2010-12-29 Koninklijke Philips Electronics N. V. Grating-based phase contrast x-ray imaging apparatus and methods
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